Non-transfected (–) and transfected (+) 293T whole cell extracts (30 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with Osteocalcin antibody (GTX132703) diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

Indirect ELISA analysis was performed by coating plate with 50 μL of recombinant full-length Osteocalcin protein at concentrations ranging from 0.938 ng/mL to 60 ng/mL. The coated protein is detected with Osteocalcin antibody (GTX132703) at 1 μg/mL.

Recombinant Osteocalcin were separated by 10% SDS-PAGE, and the membrane was blotted with Osteocalcin antibody (GTX132703) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.